A simple comet assay for archived sperm correlates DNA fragmentation to reduced hyperactivation and penetration of zona-free hamster oocytes

Chan, Philip J.; Corselli, J.; Patton, William C.; Jacobson, John D.; Chana, Steven R; King, Alan

doi:10.1016/s0015-0282(00)01655-1

Cited by 61 publications

(31 citation statements)

References 22 publications

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“…Moreover, % of DNA fragmentation was significantly negatively correlated with each of semen parameters. These findings are consistent with others (5,45,46,48,49). However, they disagree with Sepaniak et al (32); Saleh et al (40).…”

Section: Discussionsupporting

confidence: 82%

Apoptotic markers in semen of infertile men: association with cigarette smoking

El-Melegy

Ali

2011

Int. braz j urol.

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Objectives: (i) To examine the role of apoptosis in the pathogenesis of DNA damage in semen from infertile men. (ii) To assess the effects of smoking on apoptotic markers and seminal parameters among infertile men. (iii) To assess the correlation of apoptosis with conventional semen parameters. Materials and Methods:The study was carried out on 70 men with idiopathic infertility, divided into two groups: thirty infertile non smokers and forty infertile smokers. In addition to 60 fertile men (30 non smokers and 30 smokers) as control group. Each subject provided semen for analysis of parameters, determination of % of DNA fragmentation, s-Fas, caspase-3 activity levels and cotinine levels. Results:The results revealed that infertile men, particularly smokers have significantly lower semen variables and significantly higher levels of apoptotic variables (% of DNA fragmentation, s-Fas and caspase-3 activity) in addition to cotinine. Conclusions:The present findings provide additional evidence supporting the importance of the evaluation of apoptotic markers to test male infertility particularly among smokers.

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Section: Discussionsupporting

confidence: 82%

Apoptotic markers in semen of infertile men: association with cigarette smoking

El-Melegy

Ali

2011

Int. braz j urol.

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“…The clinical significance of comet assay in correlating the DNA damage with sperm concentration, morphology, mitochondrial function and oocyte peneteration has been documented in previous studies [95][96][97]. In a study by O'Connell [98] it was reported that sperm isolated from semen by density centrifugation for ART have less DNA fragmentation.…”

Section: Clinical Aspects and Relevance Of Dna Integrity Testsmentioning

confidence: 91%

Sperm DNA integrity assays: diagnostic and prognostic challenges and implications in management of infertility

Shamsi

Imam

Dada

2011

J Assist Reprod Genet

135

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Sperm is not a simple carrier of paternal genetic information but its role extends clearly beyond fertilization. Integrity of sperm genome is an essential pre-requisite for birth of healthy offspring and evaluation of sperm should entail DNA integrity analysis. DNA integrity analysis is a better diagnostic and prognostic marker of sperm reproductive potential. Conventional semen analysis emphasizes on sperm concentration, viability, motility and morphology and has been proven to be a poor indicator of reproductive potential and pregnancy outcome. To overcome the drawbacks associated with conventional semen analysis more useful fertility tests and molecular biomarkers have been explored. Among the different tests which have evolved for assessing the sperm reproductive potential, tests for sperm DNA quality are most promising. Sperm DNA damage has been closely associated with numerous indicators of reproductive health including fertilization, embryo quality, implantation, spontaneous abortion, congenital malformations and childhood diseases. It therefore has great potential as a prognostic test for both in vitro and in vivo conception. This review presents an updated account of tests that have better diagnostic and prognostic implications in the evaluation of sperm DNA damage. The basic principles, outline of methodology, advantage, disadvantage, clinical significance of each technique and implications of these tests have been discussed. The logistics of each test with respect to available resources and equipment in an andrology laboratory, the feasibility of performing these tests in routine diagnostic workup of infertile men and the opportunities and challenges provided by DNA testing in male fertility determination are also presented.

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“…Several lysis solutions including variety of chemicals have been used for this process at different time intervals in various species. [33,[36][37][38][39] The process of electrophoresis: In this process, after the incubation of revealed sperm DNA in neutral or alkaline electrophoresis solution, the revealed sperm DNA is exposed to the electrophoresis in neutral or alkaline conditions. Different electrophoresis solutions, variety of electric powers and application times were used in the electrophoresis process in previous studies.…”

Section: The Lysis Processmentioning

confidence: 99%

“…[ 34,[36][37][38][39] Assessment of comet images: In this process, the slides are incubated in the neutralization solution, air dried and stained with DNA specific stains. The stained comet images are assessed subjectively under fluorescence microscope or by computer-assisted COMET assay analysis program linked to fluorescence microscope.…”

Section: The Lysis Processmentioning

confidence: 99%

Sperm DNA and detection of DNA fragmentations in sperm

Küçük¹

2018

Turkish Journal of Urology

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The questionable effectiveness of routine sperm parameters in determining male factor infertility problems and increasing the success rates of assisted reproductive techniques have led to the investigation of more detailed sperm parameters that could affect the male fertility and reproduction. Thus, the effects of different sperm parameters such as sperm DNA integrity was started to be investigated thanks to the previously described methods such as single cell gel electrophoresis (COMET) assay, sperm chromatin structure assay (SCSA), acridine orange test (AOT), terminal deoxynucleotidyl transferase-mediated deoxyuridine (TdT) triphosphate (dUTP) nick end labeling (TUNEL) assay and sperm chromatin dispersion (SCD) test. However, studying on sperm DNA might be very complex because the sperm DNA differs from the somatic cell DNA with its unique structure. Also, the sperm DNA undergoes many changes during spermatogenesis and it is condensed by being packaged tightly with different types and numbers of protamines in different species. Despite all these difficulties, these methods provide important information about the reasons and consequences of DNA damages in sperm and the effects of these damages on reproduction.

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A simple comet assay for archived sperm correlates DNA fragmentation to reduced hyperactivation and penetration of zona-free hamster oocytes

Cited by 61 publications

References 22 publications

Apoptotic markers in semen of infertile men: association with cigarette smoking

Apoptotic markers in semen of infertile men: association with cigarette smoking

Sperm DNA integrity assays: diagnostic and prognostic challenges and implications in management of infertility

Sperm DNA and detection of DNA fragmentations in sperm

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